HLA-DRB1*07 may have a susceptibility and DRB1*04 a protective effect upon the development of a sensitization to house dust mite Dermatophagoides pteronyssinus

BACKGROUND: IgE responses to house dust mite-derived allergens seem to be the most important in the development of atopic asthma and rhinitis, but it has been difficult to demonstrate genetic control of the IgE response to the allergens. OBJECTIVE: This study was undertaken to investigate the association between sensitization to house dust mite, D. pteronyssinus (DP), and genotypes of HLA-DRB1 alleles. METHODS: DNAs were extracted from two groups of unrelated Koreans: (1) 178 with sensitization to DP; and (2) 99 age-matched non-atopic controls. Genotypes of the HLA-DRB1 alleles were determined using polymerase chain reaction (PCR)-based methods. RESULTS: The frequency of HLA-DRB1*07 was significantly increased in the DP-sensitive subjects compared with the controls (15.7% vs 4.0%, P = 0.009). Conversely, the frequencies of HLA-DRB1*04 and *14 were decreased in the DP-sensitive subjects compared with the controls (27.5% vs 45.5%, P = 0.002; 13.5% vs 24.2%, P = 0.02). Of the DRB1*04 alleles, DRB1*0403 was significantly decreased in the DP-sensitive subjects compared with the controls (3.9% vs 13.1%, P = 0.005). No significant differences were found in the distributions of the other HLA-DRB1 alleles between the two groups. CONCLUSION: HLA-DRB1*07 may have a susceptibility, and DRB1*04 and DRB1*14 may have a protective effect, upon the development of a sensitization to DP.     

T-cell sensitization to epitopes from the house dust mites Dermatophagoides pteronyssinus and Euroglyphus maynei

Background Dermatophagoides pteronyssinus and Euroglyphus maynei frequently occur in house dust but little is known about primary sensitization to the less abundant E. maynei. Objective To determine the occurrence of primary sensitization to E. maynei by T-cell responses and the crossreactivity to D. pteronyssinus. Methods The proliferative response ot peripheral blood cells to overlapping peptides from Derp I and Eur m 1 were measured as well as to peptides from Der f 1, an allergen not found in the study environment. Results The most frequent and strongest responses were to Der p 1 peptides especially in the region 105–133. However, 3/17 responders to mite peptides were stimulated predominantly by Eur m 1 peptides and a further two had their highest response to an Eur m 1 peptide. There was very little crossreactivity between Der p 1 and Eur m 1 peptides and very little response to peptides from Der f 1. Conclusion E. maynei group 1 allergens are a significant source of primary T-cell sensitization and have little T-cell crossreactivity with D. pteronyssinus or D. farinae.     

House dust mites and their allergens at selected locations in the homes of house dust mite-allergic patients

BACKGROUND: Knowledge of the occurrence of house dust mites (HDM) and their allergens in domestic locations is important when planning intervention. OBJECTIVE: The aim of this study was to describe the distribution of HDMs and their allergens before intervention in multiple locations in the homes of newly diagnosed HDM-allergic patients with a known high Der 1 concentration in their mattress dust. METHODS: Dust was collected from ten locations in the homes of eight HDM-allergic patients. Dust was analysed for allergen content with ELISAs for Der f 1, Der p 1 and Der m 1; and HDM were counted. Total allergen concentrations ( micro g Der 1/g dust) were expressed as the sum of Der f 1, Der p 1 and Der m 1. RESULTS: On mattresses the median concentration was 86 micro g Der 1/g dust (range 30-288) and 188 mites/g dust (range 12-1910). Der 1 exceeded 10 micro g/g dust in mattresses (8/8), duvets/pillows (3/8), a bedroom carpet (1/1), a living room carpet (1/6), upholstered furniture (2/8) and a curtain (1/5). Uncarpeted floors, upholstered furniture, bookshelves and walls had significantly lower Der 1 concentration than the mattresses. The relative contribution of Der p 1, Der f 1 or Der m 1 to Der 1 was related to homes, rather than to the location. Der m 1 only occurred in minute amounts. CONCLUSION: For HDM intervention, our results indicate that priority should be given to the removal of allergens from mattresses, and in addition from carpets, duvets/pillows and upholstered furniture. Dust from walls, uncarpeted floors, bookshelves and curtains appear to contribute insignificantly to the domestic HDM allergen load.     

Molecular characterization of Der p 10: a diagnostic marker for broad sensitization in house dust mite allergy

Background Tropomyosins represent clinically relevant seafood allergens but the role of mite tropomyosin, Der p 10, in house dust mite (HDM) allergy has not been studied in detail. Objective To express and purify a recombinant Der p 10 with equivalent IgE reactivity as natural Der p 10 and to evaluate its IgE reactivity and allergenic activity in HDM‐allergic patients. Methods rDer p 10 was expressed in Escherichia coli, purified and characterized by mass spectrometry and circular dichroism. It was tested for IgE reactivity in 1322 HDM‐allergic patients. Detailed IgE‐reactivity profiles to six HDM allergens (Der p 1, 2, 5, 7, 10, 21) were established for subgroups of Der p 10‐positive and ‐negative patients. The allergenic activity of rDer p 10 was evaluated in basophil degranulation experiments. Results rDer p 10 is an α‐helical protein sharing IgE epitopes with nDer p 10. It is recognized by 15.2% of HDM‐allergic patients. Der p 10‐negative patients were primarily sensitized to Der p 1 and/or Der p 2, whereas Der p 10‐positive patients reacted to several other HDM allergens besides the major allergens (Der p 1, Der p 2) or showed a rather selective Der p 10 reactivity. The allergenic activity of Der p 10 was generally low but patients could be identified who suffered from clinically relevant HDM allergy due to Der p 10 sensitization. Conclusion and Clinical Relevance Der p 10 may be a diagnostic marker for HDM‐allergic patients with additional sensitization to allergens other than Der p 1 and Der p 2. Such patients may require attention when allergen‐specific immunotherapy is considered. Cite this as: Y. Resch, M. Weghofer, S. Seiberler, F. Horak, S. Scheiblhofer, B. Linhart, I. Swoboda, W. R. Thomas, J. Thalhamer, R. Valenta and S. Vrtala, Clinical & Experimental Allergy, 2011 (41) 1468–1477.     

House-dust mite content in mattresses in relation to residential characteristics and symptoms in atopic and nonatopic children living in northern Norway

The occurrence of house-dust mites (HDMs) was investigated in the mattresses of 19 children previously found to be skin prick test (SPT) positive to HDM and in 19 nonatopic children derived from an extensive survey of 424 schoolchildren, all living in northern Norway. Domestic mites were counted and identified microscopically. Mite counts ranging from 10 to 1800 mites per gram mattress dust were found in 10 of the 19 HDM-sensitized children compared to none in the control group, corresponding to an odds ratio of more than 20. Of the 540 domestic mites found, 70 were identified by species. Dermatophagoides pteronyssinus (Dpt) was the only HDM species identified (64 mites), while five were storage mites and one was a Tarsonemus species. Positive radioallergosorbent tests (RAST) to Dpt were demonstrated in 9/10 children with and in 5/9 without mite infestation compared to none in the control group. Elevated IgE levels were also found more frequently in children with mite-infested mattresses than in those without. IgE levels were within normal levels in all 19 children in the control group. Latent atopy was found in four children, three with and one without mite infestation. There was no correlation between the concentration of mites and the degree of sensitization. Poor ventilation, increased humidity, and water leak(s) were associated with the presence of domestic mites in mattresses. As HDM growth is highly dependent on humidity and microhabitat, it should be possible to avoid HDM exposure and allergy in this region.     

哮喘患儿家庭内尘螨过敏原分布特征及其临床意义

目的了解北京地区尘螨过敏性哮喘患儿家庭环境内尘螨过敏原含量分布特征,初步探讨尘螨过敏原暴露水平的临床意义。方法选取54例尘螨过敏性哮喘患儿,其中男性37例,女性17例;年龄3～16岁,平均年龄8岁2个月。采集患儿家庭中床垫、枕头、卧室地板、客厅地板及沙发的灰尘,采用酶联免疫吸附分析（ELISA）测定以上灰尘样本中户尘螨1组过敏原（Der p1）和粉尘螨1组过敏原（Der f1）的含量;应用荧光ELISA测定患儿血清尘螨特异性IgE浓度;评估患儿哮喘临床控制情况,应用化学发光法测定患儿呼出气一氧化氮浓度（FeNO）。结果采集灰尘样本255份,以中位数（最小值～最大值）表示尘螨过敏原含量,床垫、枕头和沙发灰尘样本中Derf1和Derp1的含量显著高于卧室地板和客厅地板灰尘样本中尘螨过敏原含量。Derf1平均含量为0.13μg/g,显著高于Derp1平均含量0.02μg/g（P〈0.05）。Derp1和Derf1联合暴露的最高含量平均为2.18（0.07～54.59）μg/g。Der p1和Der f1联合最高暴露含量≥10.00μg/g、2.00～10.00μg/g、0.05～2.00μg/g的例数分别为4例（7.4%）、24例（44.4%）、26例（48.1%）。其中未控制组患儿家庭内尘螨过敏原最高暴露水平为27.41（0.23～54.59）μg/g,均显著高于部分控制组和控制组哮喘患儿尘螨过敏原最高暴露水平1.66（0.07～26.27）μg/g、2.90（0.37～33.75）μg/g（P〈0.05）。不同sIgE浓度分级组间尘螨过敏原最高暴露水平的差异、不同FeNO浓度范围组间尘螨过敏原最高暴露水平差异均无统计学意义。结论北京地区尘螨过敏性哮喘患儿家庭尘螨以Der f1为主,床垫、枕头及沙发灰尘样本是Der p1和Der f1的主要来源;哮喘未控制者的尘螨过敏原最高暴露水平明显增高。     

Mite (Der p 1, Der f 1) and cat (Fel d 1) allergens in the homes of babies with a family history of allergy

Carpet and floor dust samples were collected in four different seasons, from 39 Swedish homes of babies with a family history of allergy. House-dust mite ( Der p 1, Der f 1 ) and cat ( Fel d 1 ) allergen contents were determined by mab ELISA, and the levels were related to various environmental factors. Both mite and cat antigens were detected in 94% of the samples and in all homes, but the levels were low ( Der p 1 , range 15 ng–1944 ng/g fine dust; Der f 1 , range 14 ng–264 ng/g of fine dust; Fel d 1 , range 16 ng–3120 ng/g fine dust). Mite-allergen levels were significantly higher ( P <0.001) in floor dust than in carpets, and D. pteronyssinus predominated. In contrast, the levels of cat antigen were significantly ( P <0.05) higher in carpets than in floor dust. There was no clear relation between mite-allergen levels and type of house, except that the higher values were found in homes with dampness problems. Cat-allergen levels were higher than total mite-allergen content, and the highest levels were found in homes with a cat ( P <0.05). Rather high concentrations of cat allergen were also found in homes without a cat, which may explain why cat sensitization is so common in Sweden. As the prevalence of house-dust mite sensitivity is increasing in Swedish children, and as the individual patient threshold for eliciting symptoms varies, we suggest that sensitization may possibly occur at a lower exposure level than generally accepted as risk level for sensitization (2 μg/g dust).     

Absence of continuous epitopes in the house dust mite major allergens Der p I from Dermatophagoides pteronyssinus and Der f I from Dermatophagoides farinae

Background The house dust mite has been shown to be an important source of domestic allergens associated with immediate hypersensitivities. The Group I mite allergens Der p I from Dermatophagoides pteronyssinus and Der f I from D. farinae display extensive amino acid sequence homology and have similarities with cysteine protease enzymes.
Objective The availability of the complete amino acid sequences for these allergens allowed us to search for the allergic detertninants within these molecules. The aim of the present investigation was to identify any continuous IgE-binding epitopes within these amino acid sequences. We also sought to test the validity of previously reported Der p I peptide epitope sequences.
Methods In order to identity any continuous IgE epitopes, the amino acid sequences of Der p I and Der f I were synthesized as decapeptides overlapping in sequence and coupled to plastic pins. The specific IgE-binding capacity of these peptides was assayed using an enzyme-linked biotin-streptavidin procedure and sera from patients known to be sensitive to these allergens. Previously reported Der p I peptide epitopes were synthesized as free peptides and tested for their ability to inhibit specific IgE binding to allergen extract discs.
Results None of the pin-coupled Der p I or Der f I peptides was found by the continuous epitope mapping procedure to bind significantly to specific IgE in the sera of hypersensitive patients. The previously reported Der p I peptide epitopes did not inhibit specific IgE binding to mite extract discs.
Conclusion The specific IgE binding epitopes of the house dust mite allergens Der p I and Der f I are discontinuous in nature.     

Silverfish protein in house dust in relation to mite and total arthropod level

Two assays have been developed to measure arthropod levels in house dust. The first assay measures silverfish antigens. The second assay measures invertebrate tropo-myosin and gives a global assessment of the level of atthropod-derived material. These assays and a Der p 1 and Der p 2 assay were used to analyse 53 dust samples. In most dust samples the ratio of tropomyosin/Der p 2 was higher than in mite body extract, indicating that the assay measures other arthropods besides mites. Silverfish antigen was detectable in most of the dust samples. In many homes in which the inhabitants were unaware of the presence of silverfish, silverfish antigen was detectable. Therefore for information on exposure an immunochemical analysis is superior to a questionnaire.     

Monoclonal antibodies to recombinant Der f 2 and development of a two-site ELISA sensitive to major Der f 2 isoallergen in Korea

BACKGROUND: Der f 2 is a major sensitizing allergen in patients allergic to house dust mites worldwide. Isoforms of Der f 2 have been reported and are known to have different antigenicities. The aim of this study was to facilitate antigenic analysis and to develop an improved method for the detection of Der f 2 isoallergen, which is prevalent in Korea. METHODS: A two-site ELISA was developed with monoclonal antibodies (mAbs) which were produced against recombinant Der f 2 (rDer f 2) and applied to assess Der f 2 in bedding samples. RESULTS: A major isoform of Der f 2, found in Korea, was found to have amino acid variations especially at position 100 from lysine to glutamic acid, which is known to reduce significantly the binding affinity of mAbs when used to assess group 2 allergens. The detection limit of the developed two-site ELISA was determined to be about 8 ng/ml with rDer f 2 and 1 microg/ml with Derntatophagoides farinae crude extract. The average amount of Der f 2 in dust obtained from bedding samples from 89 homes in Seoul was estimated to be 25.61+/-10.70 microg/g dust. CONCLUSIONS: Assays using mAbs for rDer f 2 could be useful for the assessment of environmental allergen exposure and mAbs could be used to further characterize the isoallergens of Der f 2.     

Sensitization to house dust mites in Reykjavik, Iceland, in the absence of domestic exposure to mites

BACKGROUND: House dust mites are common sources of indoor allergens. In Reykjavik, Iceland, 9% of the young adult population had serum-specific IgE to Dermatophagoides pteronyssinus. Sensitization to mites is usually assumed to be due to exposure to house dust mites in the indoor environment. This investigation was carried out to measure the concentrations of house dust mite allergens and to investigate which species of mites were present in beds in Iceland. METHODS: A total of 197 randomly selected adults were visited at home using the European Community Respiratory Health Survey (ECRHS) II Indoor protocol. Dust samples were collected from mattresses for measurement of house dust mite allergen concentrations and to estimate the number and type of house dust mites. Additional samples from mattresses and floors were collected from the homes of 10 patients with positive skin prick tests (SPT) to D. pteronyssinus. House dust mite allergen concentrations were measured using ELISA and examination of mite species was carried out using microscopy. Climatic parameters were assessed using psychrometer readings in the bedrooms and outdoors. RESULTS: We found two single mite specimens, both D. pteronyssinus, in two dust samples. Mite allergen analyses indicated that two other dust samples had Der f 1 results close to the cut-off of 0.1 microg/g of dust. No samples were positive for Der p 1. In an additional collection of dust from the homes of 10 SPT-positive patients no Dermatophagoides spp. were found. CONCLUSIONS: Reykjavik citizens are exposed to extremely low amounts of house dust mite allergens in their homes. Possible alternative sources for sensitization are discussed, such as bird nests, exposure from travelling abroad, or other mites or invertebrates that cross-react with house dust mite allergens. Our findings suggest that exposures other than to house dust mites indoors are possible sources of mite allergen exposure.     

Skin tests, T cell responses and self-reported symptoms in children with allergic rhinitis and asthma due to house dust mite allergy

Background In allergic responses, a distinction is made between an early-phase response, several minutes after allergen exposure, and a late-phase response after several hours. During the late phase, eosinophils and T cells infiltrate the mucosa and play an important role in inflammation. Objective The aim of this study was to examine the relationship between allergen-induced late-phase skin responses and in vitro T cell reactivity. In addition, the relationship between allergen-induced skin or T cell responses and the severity of self-reported symptoms was studied in children with house dust mite allergy. Methods A total of 59 house dust mite-allergic children (6–18 years) were recruited in general practice. These children or their parents rated their nasal and asthma symptoms on diary cards during 1 month. Allergen skin tests were performed and read after 15 min (early phase) and 6 h (late phase). Allergen-specific T cell proliferation was determined, and Th2 cytokine (IL-5 and IL-13) secretion was analysed. Results The size of the late-phase skin response correlated with in vitro T cell proliferation (r_s=0.38, P=0.003) but not with Th2 cytokine secretion (r_s=0.16, P=0.2 for both IL-5 and IL-13). Moreover, the late-phase skin response and T cell proliferation correlated with asthma symptoms (r_s=0.30, P=0.02 for skin response and r_s=0.28, P=0.03 for T cell proliferation) but not with nasal symptoms (r_s=0.19, P=0.15 for skin response and r_s=0.09, P=0.52 for T cell proliferation). The early-phase skin response correlated with the nasal symptom score (r_s=0.34, P=0.01) but not with asthma symptom scores (r_s<0.005, P=0.97). Conclusion In this study, the late-phase skin test response correlated with in vitro T cell proliferation but not with Th2 cytokine secretion. We found weak or no correlations between late-phase skin responses and symptoms of asthma or rhinitis in children with house dust mite allergy. This suggests that late-phase skin responses reflect certain T cell properties but are of limited value for the evaluation of airway symptoms in atopic children.     

Mite allergens in relation to home conditions and sensitization of asthmatic children from three climatic regions

We investigated the levels of mite (Der p I and Der f I) allergen in dust from bedrooms, living rooms, kitchens, and bathrooms from 130 homes of asthmatic children in three climatic zones of Sweden. Bedroom dust samples included the child's mattress, carpets, floors, and other plain surfaces. Living-room dust samples were taken from sofas and other furniture, carpets, floors, and other plain surfaces. The allergen levels were related to home characteristics, including absolute indoor humidity (AIH), relative humidity (RH), and air changes per hour (ach). Mite allergen was detected in 62% of the homes. Levels of Der p I varied between <16 ng and 50 μg/g dust, and Der f I between ≤16 ng and 73 μg/g dust. Because we have designed a composite type of dust collection in our study, the allergen levels found tend to average down the results. Mite allergen levels were higher in homes with dampness problems, in homes with a smoker, and in homes without a basement. Homes with high absolute humidity (7 g/kg) or relative humidity (45%) and poor ventilation (≤0.5 ach) contained higher levels of mite allergens than homes with lower humidity and better ventilation. However, the number of ach measurements in homes was not high, and few homes had ≤0.5 ach. Sensitization to house-dust mites was more common in southern than in northern and central Sweden. High levels of house-dust mite allergen in a temperate climate where mites are not ubiquitous are thus associated with dampness problems in homes and with tobacco smoking. Our data confirm and extend previous findings that high AIH and RH and poor ventilation increase the risk of mite infestation in homes. It seems to be important and necessary to control indoor humidity and ventilation levels, to avoid high mite allergen exposure in a temperate climate, because 34% of mite-sensitized asthmatic children were exposed to levels of mite allergen >2 μg/g dust in their homes. The study also shows that mite allergen levels below the suggested threshold level (2 μg/g dust) are associated with mite sensitivity in children with perennial symptoms of asthma.     

Seasonal differences in airway hyperresponsiveness in asthmatic patients: relationship with allergen exposure and sensitization to house dust mites

Background The degree of airway hyperresponsiveness in allergic asthmatic patients may be influenced by changes in environmental exposure to inhalant allergens. Objective This study investigates the relationship between seasonal changes in exposure to house dust mite (HDM) allergens and non-specific airway hyperresponsiveness in asthmatic patients with multiple sensitizations to inhaled allergens. Methods In 43 asthmatic patients sensitized to several inhalant allergens, lung function (FEV₁), airway hyperresponsiveness (PC₂₀ histamine), serum total IgE, house dust mite (HDM) specific IgE and number of peripheral blood eosinophils were measured during autumn 1990 (September-November) and spring 1991 (March—May). During each season. floor dust samples were collected twice from living- and bedrooms and the concentration of the HDM allergens Der p 1 and Der p 2 determined. Results More severe airway hyperresponsiveness (lower PC₂₀ histamine) during autumn was only found in patients sensitized to HDM(n= 32; autumn: 2.05mg/mL, spring: 4.51mg/mL (geometric means), P <0.0 1), whereas in patients not sensitized to HDM (n= 11) similar values were observed in both seasons (3.44 and 4.52 mg/mL. respectively, P= 0.56). More severe airway hyperresponsiveness of HDM sensitized patients in autumn was significantly associated with higher Der p 1 concentrations in floor dust. Aside from airway hyperresponsiveness, seasonal changes in serum total IgE and number of peripheral blood eosinophils were seen in patients sensitized to HDM, Conclusions In allergic asthmatic patients, airway hyperresponsiveness may increase during autumn, depending on sensitization to HDM and an increase of exposure to HDM allergen.     

Usefulness of specific immunotherapy in patients with atopic dermatitis and allergic sensitization to house dust mites: a multi-centre, randomized, dose-response study

BACKGROUND: The effect of specific immunotherapy (SIT) on eczema in atopic dermatitis is not known. Therefore, a multi-centre, randomized dose-response trial, double-blind with respect to the efficacy of a biologically standardized depot house dust mite preparation was performed. METHODS: Eighty-nine adults with a chronic course of atopic dermatitis, SCORAD >or=40 and allergic sensitization to house dust mites [CAP-FEIA >or=3] were included, of whom 51 completed the study. Subcutaneous SIT with a house dust mite preparation (Dermatophagoides pteronyssinus/D. farinae) applying maintenance doses of 20, 2,000 and 20,000 SQ-U in weekly intervals for 1 year. The main outcome measures addressed the change of the SCORAD as average of the values after 9 and 12 months of SIT in comparison with the value at baseline. RESULTS: The SCORAD declined in the three dose groups in a dose-dependent manner (P = 0.0368, Jonckheere-Terpstra test) and was significantly lower in the two high-dose groups (2,000, 20,000 SQ-U) compared with the low-dose group of 20 SQ-U (P = 0.0379, U-test) after 1 year of SIT. The use of topical corticosteroids was significantly reduced with higher doses (P = 0.0007, Mantel-Haenszel chi-square test). CONCLUSIONS: Allergen-SIT for 1 year with a house dust mite preparation is able to improve the eczema in patients with atopic dermatitis who are sensitized to house dust mite allergens and reduces the need for topical corticosteroids. SIT may be valuable in the treatment of this chronic skin disease.     

Asthma and allergy in Russian and Norwegian schoolchildren: results from two questionnaire-based studies in the Kola Peninsula, Russia, and northern Norway

BACKGROUND: Previous studies have shown that the prevalence of asthma and allergy in children is lower in Eastern than Western Europe. METHODS: We have compared the prevalence of asthma, respiratory symptoms, allergic rhinoconjunctivitis, and atopic dermatitis in schoolchildren aged 7-13 years in a questionnaire-based study conducted in the city of Nikel on the Kola Peninsula, Russia, in 1994 (n = 1143) and another conducted in northern Norway in 1995 (n = 8676). RESULTS: The prevalence of diagnosed asthma was 5.1% in Russian children and 8.6% in Norwegian children; RR =0.58 (95% CI: 0.44-0.76). The prevalence of all respiratory symptoms was higher in Russian children. The prevalence of allergic rhinoconjunctivitis was 16.9%, in Russian children and 22.1%, in Norwegian children: RR =0.74 (95% CI: 0.65-0.85). The prevalence of atopic dermatitis was 7.4% in Russian children and 19.7% in Norwegian children; RR=0.38 (95% CI: 0.31-0.46). CONCLUSIONS: We conclude that the prevalence of diagnosed asthma, allergic rhinoconjunctivitis, and atopic dermatitis was higher in Norwegian than Russian schoolchildren. The higher prevalence of respiratory symptoms in Russian children probably reflects a higher prevalence of undiagnosed, nonallergic asthma.     

Asthma and allergic symptoms in relation to house dust endotoxin: Phase Two of the International Study on Asthma and Allergies in Childhood (ISAAC II)

Background Several studies have consistently reported inverse associations between exposure to endotoxin in house dust and atopy. With regard to the association between house dust endotoxin and asthma, the results are inconsistent. Objectives To study the association between house dust endotoxin levels and respiratory symptoms and atopy in populations from largely different countries. Methods Data were collected within the International Study on Asthma and Allergies in Childhood Phase Two, a multi‐centre cross‐sectional study of 840 children aged 9–12 years from six centres in the five countries of Albania, Italy, New Zealand, Sweden and the United Kingdom. Living room floor dust was collected and analysed for endotoxin. Health end‐points and demographics were assessed by standardized questionnaires. Atopy was assessed by measurements of allergen‐specific IgE against a panel of inhalant allergens. Associations between house dust endotoxin and health outcomes were analysed by logistic regression. Odds ratios (ORs) were presented for an overall interquartile range increase in exposure. Results Many associations between house dust endotoxin in living room floor dust and health outcomes varied between countries. Combined across countries, endotoxin levels were inversely associated with asthma ever [adjusted OR (95% confidence interval (CI)) 0.53 (0.29–0.96) for endotoxin levels per m² of living room floor] and current wheeze [adjusted OR (95% CI) 0.77 (0.64–0.93) for endotoxin levels per gram of living room floor dust]. There were inverse associations between endotoxin concentrations and atopy, which were statistically significant in unadjusted analyses, but not after adjustment for gender, parental allergies, cat and house dust mite allergens. No associations were found with dust quantity and between endotoxin exposure and hayfever. Conclusion These findings suggest an inverse association between endotoxin levels in living room floor dust and asthma in children.